端粒逆转录酶基因修饰对绵羊骨髓间充质干细胞端粒酶活性影响的研究

Study on the Effect of TERTGene Modification on Activity of Telomerase in Bone Marrow Mesenchymal Stem Cells of Ovis Aries

骨髓间充质干细胞(BMSC)具有多向分化的特性,但其生命周期有限,为进一步探讨端粒逆转录酶(TERT)对端粒酶活性的影响,本试验利用将外源性TERT导入间充质干细胞技术,构建重组表达载体pcDNA3.1-EGFP-TERT,将其转染BMSC,通过筛选及鉴定,结果显示成功构建了重组表达载体pcDNA3.1-EGFP-TERT。将TERT-BMSCs在体外进行传代培养及生长曲线的测定,利用PCR技术对其原有干细胞因子的表达情况进行了鉴定。结果显示,TERT-BMSC具有快速扩增的能力,细胞形态良好,且仍具有干细胞特性。本试验结果揭示了外源性TERT基因的表达能激活绵羊BMSC的端粒酶活性。

Bone marrow mesenchymal stem cells （BMSC） had multiple differentiation potentiality, but their life cycle was limited,in order to make further study on the effect of telomerase reverse transcriptase （TERT） gene on activity of telomerase,our research adopted the method of trans- fecting exogenous TERT into mesenchymal stem cell technology, constructed recombinant ex- pressing vector pcDNA3. 1-EGFP-TERT, and then transfecting into ovine BMSCs, and through selection and identification,recombinant expressing vector pcDNA3.1-EGFP-TERT was success- fully expressed in BMSCs. Then the TERT-BMSCs were cultured in vitro and the determination of growth curve was conducted, finally,the expression of the original stem cell factor were identi- fied by PCR technique. The results showed that TERT-BMSC had the ability of rapid amplifica- tion, cell shape was good and still had the characteristics of stem cells. We got the conclusion that the expression of exogenous TERT gene could activate the activity of BMSCs~ telomerase.